Morampudi Vijay, Bhinder Ganive, Wu Xiujuan, Dai Chuanbin, Sham Ho Pan, Vallance Bruce A, Jacobson Kevan
Division of Gastroenterology, BC Children's Hospital.
Division of Gastroenterology, BC Children's Hospital;
J Vis Exp. 2014 Feb 27(84):e51297. doi: 10.3791/51297.
Inflammatory Bowel Diseases (IBD), including Crohn's Disease and Ulcerative Colitis, have long been associated with a genetic basis, and more recently host immune responses to microbial and environmental agents. Dinitrobenzene sulfonic acid (DNBS)-induced colitis allows one to study the pathogenesis of IBD associated environmental triggers such as stress and diet, the effects of potential therapies, and the mechanisms underlying intestinal inflammation and mucosal injury. In this paper, we investigated the effects of dietary n-3 and n-6 fatty acids on the colonic mucosal inflammatory response to DNBS-induced colitis in rats. All rats were fed identical diets with the exception of different types of fatty acids [safflower oil (SO), canola oil (CO), or fish oil (FO)] for three weeks prior to exposure to intrarectal DNBS. Control rats given intrarectal ethanol continued gaining weight over the 5 day study, whereas, DNBS-treated rats fed lipid diets all lost weight with FO and CO fed rats demonstrating significant weight loss by 48 hr and rats fed SO by 72 hr. Weight gain resumed after 72 hr post DNBS, and by 5 days post DNBS, the FO group had a higher body weight than SO or CO groups. Colonic sections collected 5 days post DNBS-treatment showed focal ulceration, crypt destruction, goblet cell depletion, and mucosal infiltration of both acute and chronic inflammatory cells that differed in severity among diet groups. The SO fed group showed the most severe damage followed by the CO, and FO fed groups that showed the mildest degree of tissue injury. Similarly, colonic myeloperoxidase (MPO) activity, a marker of neutrophil activity was significantly higher in SO followed by CO fed rats, with FO fed rats having significantly lower MPO activity. These results demonstrate the use of DNBS-induced colitis, as outlined in this protocol, to determine the impact of diet in the pathogenesis of IBD.
炎症性肠病(IBD),包括克罗恩病和溃疡性结肠炎,长期以来一直被认为与遗传基础有关,最近又发现与宿主对微生物和环境因素的免疫反应有关。二硝基苯磺酸(DNBS)诱导的结肠炎使人们能够研究与IBD相关的环境触发因素(如压力和饮食)的发病机制、潜在治疗方法的效果以及肠道炎症和黏膜损伤的潜在机制。在本文中,我们研究了膳食n-3和n-6脂肪酸对大鼠结肠黏膜对DNBS诱导的结肠炎的炎症反应的影响。在直肠内给予DNBS之前,所有大鼠均喂食相同的饮食,只是脂肪酸类型不同(红花油(SO)、菜籽油(CO)或鱼油(FO)),持续三周。给予直肠内乙醇的对照大鼠在为期5天的研究中体重持续增加,而喂食脂质饮食的DNBS处理大鼠体重均下降,喂食FO和CO的大鼠在48小时时体重显著下降,喂食SO的大鼠在72小时时体重显著下降。DNBS处理后72小时体重恢复增加,到DNBS处理后5天,FO组的体重高于SO或CO组。DNBS处理5天后收集的结肠切片显示有局灶性溃疡、隐窝破坏、杯状细胞减少以及急性和慢性炎症细胞的黏膜浸润,不同饮食组的严重程度不同。喂食SO的组损伤最严重,其次是CO组,喂食FO的组组织损伤程度最轻。同样,结肠髓过氧化物酶(MPO)活性是中性粒细胞活性的标志物,在喂食SO的大鼠中显著高于喂食CO的大鼠,喂食FO的大鼠MPO活性显著较低。这些结果表明,按照本方案所述使用DNBS诱导的结肠炎来确定饮食在IBD发病机制中的影响。